P62-ZZ small molecule modulators

ABSTRACT

A compound, or a pharmaceutically acceptable salt thereof, having a formula I of: 
     
       
         
         
             
             
         
       
         
         
           
             wherein Ar is an arylene or heteroarylene; 
           
         
         R 1  has a structure of: 
       
    
     
       
         
         
             
             
         
       
         
         
           
             wherein W is an alkanediyl, alkenediyl, a carbonyl, or a combination thereof; 
             X is —NR 5 —, wherein R 5  is H or an alkyl, or —O—; and 
             Y is optionally-substituted cycloalkyl, optionally-substituted cycloalkyl-substituted alkyl, optionally-substituted heterocycloalkyl, or optionally-substituted heterocycloalkyl-substituted alkyl; 
           
         
         each R 2  is the same or different and has a structure of: 
       
    
     
       
         
         
             
             
         
       
         
         
           
             wherein Z is —NR 6 —, wherein R 6  is H or an alkyl, —O—, —S—, or —CH 2 —; 
             Z 1  is (—CH 2 —) m  wherein m is 0 to 5, or an alkenediyl having 2 to 6 carbon atoms; 
             Cy is a 3-8-membered cycloalkyl, heterocycloalkyl, aryl or heteroaryl ring; and 
             each R 4  is the same or different and is selected from hydroxy, halogen, substituted or unsubstituted alkoxy, substituted or unsubstituted alkyl, or amino; and c is 0 to 5; and 
             each R 3  is the same or different and is selected from hydroxy, halogen, substituted or unsubstituted alkoxy, substituted or unsubstituted alkyl, amino, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, or a nitro, 
             wherein a is 2 to 5, and b is 0 to 3.

CROSS REFERENCE TO RELATED APPLICATIONS

This application is the U.S. National Stage of International ApplicationNo. PCT/US2016/036241, filed Jun. 7, 2016, which was published inEnglish under PCT Article 21(2), which in turn claims the benefit ofU.S. Provisional Application No. 62/174,465, filed Jun. 11, 2015, whichis incorporated herein by reference.

BACKGROUND

Sequestosome-1 (SQSTM1/p62) is rich with protein-interacting domains,including an N-terminal PB1 domain, a ZZ-type zinc finger domain, aTRAF6-binding domain (TBS), the LC3-interacting region (LIR), theKEAP1-interacting region (KIR), and a C-terminal ubiquitin bindingdomain (UBA). While p62 may primarily act as a key adaptor for thedegradation of protein aggregates, cytoplasmic bodies and malfunctioningorganelles by selective autophagy, it continues to gain interest for itsintimate and complex involvement in a number of cell signaling pathwaysand functions. Generally, p62 has been shown to play important roles inprotein ubiquitination, triggering cell autophagy and apoptosis intumorigenesis, and it is particularly implicated in the activation ofthe transcription factor NF-κB, p38MAPK and mTOR pathways that helpregulate cell homeostasis.

Studies have revealed that p62 takes part in selective autophagy.Autophagy is a tightly regulated conserved catabolic process occurringin all cells and thus has important implications for cell homeostasis,development and immune response among other functions. For example,microtubule-binding tau proteins in neurons are transferred toproteasomes by p62. Interestingly, disruption of the p62 gene in miceresults in a phenotype resembling Alzheimer's disease. Similarly,accumulations of aggregated mutant huntingtin protein, the pathologicalbasis of Huntington's disease, have been found to contain p62 anddeletion of its UBA domain increases cell death by mutant huntingtin.p62 has also been found in cytoplasmic aggregates described inParkinson's disease and amyotrophic lateral sclerosis as well as inbreast cancer tumors. Additionally, targeted deletion of mice p62 hasbeen found to lead to insulin and leptin resistance, type 2 diabetes andobesity. As emerging evidence supports p62's implication in severalvarying diseases, thus targeting Sequestosome-1 (SQSTM1/p62) is highlysignificant for drug design and discovery.

It is known that p62 plays a key role in autophagy and cell signalinginvolving the NF-kB, p38MAPK and mTOR pathways. In particular, studieswith p62-deficient mice have demonstrated that one p62 function is tocontrol osteoclastogenesis and bone remodeling. Normal osteoclastfunction relies on this regulation of the NF-kB pathway by p62. The Zdomain is implicated in this mechanism, as it binds to RIP1 protein,which contains a “death” domain that interacts with TNF receptor, whichcan then activate NF-kB and p38MAPK signaling. In light of these andprevious findings, p62 is an attractive drug target for a myriad ofdiseases, particularly multiple myeloma (MM) and related cancers as wellas other diseases such as neurodisorder and diabetics diseases.

Among the diseases mentioned above, MM is an incurable hematologicmalignancy, characterized by the dysregulated proliferation of plasmacells and progressive bone destruction in up to 80% patients. Despitethe introduction of novel and more potent treatment regimens includingthalidomide and bortezomib, MM is still the second most prevalenthematological malignancy. As reported by the Leukemia & Lymphoma Society(Facts 2009-2010), both the MM patient number and new diagnosed caseshave markedly increased each year. Therefore, novel therapeutics thateffectively inhibits tumor growth and overcome conventional drugresistance are urgently needed.

SUMMARY

Disclosed herein are compounds, or pharmaceutically acceptable saltsthereof, having a formula I of:

-   -   wherein Ar is an arylene or heteroarylene;        R¹ has a structure of:

-   -   wherein W is an alkanediyl, alkenediyl, a carbonyl, or a        combination thereof;    -   X is —NR⁵—, wherein R⁵ is H or an alkyl, or —O—; and    -   Y is an optionally-substituted cycloalkyl,        optionally-substituted cycloalkyl-substituted alkyl;    -   optionally-substituted heterocycloalkyl, or an        optionally-substituted heterocycloalkyl-substituted alkyl;        each R² is the same or different and has a structure of:

-   -   wherein Z is —NR⁶—, wherein R⁶ is H or an alkyl, —O—, —S—, or        —CH₂—;    -   Z¹ is (—CH₂—)_(m) wherein m is 0 to 5, or an alkenediyl having 2        to 6 carbon atoms;    -   Cy is a 3-8-membered cycloalkyl, heterocycloalkyl, aryl or        heteroaryl ring; and    -   each R⁴ is the same or different and is selected from hydroxy,        halogen, substituted or unsubstituted alkoxy, substituted or        unsubstituted alkyl, or amino; and c is 0 to 5; and    -   each R³ is the same or different and is selected from hydroxy,        halogen, substituted or unsubstituted alkoxy, substituted or        unsubstituted alkyl, amino, substituted or unsubstituted        cycloalkyl, substituted or unsubstituted heterocycloalkyl,        substituted or unsubstituted aryl, or a nitro, wherein a is 2 to        5, and b is 0 to 3.

Also disclosed herein is a method for treating a p62-mediated disease ina subject, the method comprising administering to a subject in needthereof a therapeutically effective amount of at least one p62-ZZinhibitor compound disclosed herein.

Further disclosed herein is a method of modulating p62 activity instromal cells, comprising contacting stromal cells with at least onep62-ZZ inhibitor compound disclosed herein.

Additionally disclosed herein is a method of inhibiting multiple myelomacell growth, comprising contacting multiple myeloma cells with at leastone p62-ZZ inhibitor compound disclosed herein.

The foregoing will become more apparent from the following detaileddescription, which proceeds with reference to the accompanying figures.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is a schematic showing various signaling pathways.

FIG. 2 is a schematic showing in vitro biological assays to validatescreened compounds for multiple myeloma cells growth via inhibition ofp62-zz domain and PKCC phosphorylation.

FIG. 3 is a graph showing the antitumor activity of compound XIELP1-106in the treatment of a RPMI-8226 human multiple myeloma xenograft model.

FIG. 4 is a graph showing the survival curves of subcutaneous RPMI822tumor-bearing mice.

DETAILED DESCRIPTION Overview

Described herein are new p62-ZZ inhibitors that affect, for example,stromal cells and multiple myeloma cells. The novel p62-ZZ inhibitorsmay demonstrate micromolar (˜2 μM) inhibition activity against multiplemyeloma cell growth without toxicity to normal stromal cells. In certainembodiments, the compounds are selective p62-ZZ inhibitors meaning thatthe compounds exhibit inhibition activity that is selective for the ZZdomain relative to other p62 domains.

Sequestosome 1 (p62) plays a key role in the formation of signalingcomplexes that result in NF-KB, p38 MAPK and PI3K activation in themarrow microenvironment of patients with MM. In contrast to treatingsubjects with inhibitors of each of the multiple signaling pathwaysactivated in marrow stromal cells by MM cells (e.g., NF-KB or p38 MAPK),blocking the function of p62 should inhibit the activation of themultiple pathways mediated by p62 and have a broader effect on the bonemarrow microenvironment.

Terminology

The following explanations of terms and methods are provided to betterdescribe the present compounds, compositions and methods, and to guidethose of ordinary skill in the art in the practice of the presentdisclosure. It is also to be understood that the terminology used in thedisclosure is for the purpose of describing particular embodiments andexamples only and is not intended to be limiting.

As used herein, the singular terms “a,” “an,” and “the” include pluralreferents unless context clearly indicates otherwise. Also, as usedherein, the term “comprises” means “includes.”

“Administration of” and “administering a” compound should be understoodto mean providing a compound, a prodrug of a compound, or apharmaceutical composition as described herein. The compound orcomposition can be administered by another person to the subject (e.g.,intravenously) or it can be self-administered by the subject (e.g.,tablets).

“Alkanediyl” refers to a divalent group of the general formula—C_(n)H_(2n)— derived from aliphatic, hydrocarbons. A lower alkanediylhas 1 (also referred to as a methylene radical) to 10 carbon atoms, moreparticularly 1 to 5 carbon atoms.

“Alkenediyl” refers to a divalent group formed from alkanes by removalof two hydrogen atoms from the same carbon atom, the free valencies ofwhich are part of a double bond. A lower alkenediyl has 2 to 10 carbonatoms, more particularly 1 to 5 carbon atoms.

The term “alkenyl” refers to a hydrocarbon group of 2 to 24 carbon atomsand structural formula containing at least one carbon-carbon doublebond. A “lower alkenyl” group has 1 to 10 carbon atoms.

The term “alkyl” refers to a branched or unbranched saturatedhydrocarbon group of 1 to 24 carbon atoms, such as methyl, ethyl,n-propyl, isopropyl, n-butyl, isobutyl, t-butyl, pentyl, hexyl, heptyl,octyl, decyl, tetradecyl, hexadecyl, eicosyl, tetracosyl and the like. A“lower alkyl” group is a saturated branched or unbranched hydrocarbonhaving from 1 to 10 carbon atoms. Preferred alkyl groups have 1 to 4carbon atoms. Alkyl groups may be “substituted alkyls” wherein one ormore hydrogen atoms are substituted with a substituent such as halogen,cycloalkyl, alkoxy, amino, hydroxyl, aryl, or carboxyl.

The term “alkylaryl” refers to a group in which an alkyl group issubstituted for a hydrogen atom of an aryl group. An example is —Ar—R,wherein Ar is an arylene group and R is an alkyl group.

The term “alkoxy” refers to a straight, branched or cyclic hydrocarbonconfiguration and combinations thereof, including from 1 to 20 carbonatoms, preferably from 1 to 8 carbon atoms, more preferably from 1 to 4carbon atoms that include an oxygen atom at the point of attachment. Anexample of an “alkoxy group” is represented by the formula —OR, where Rcan be an alkyl group, optionally substituted with an alkenyl, alkynyl,aryl, aralkyl, cycloalkyl, halogenated alkyl, or heterocycloalkyl groupas described above. Suitable alkoxy groups include methoxy, ethoxy,n-propoxy, i-propoxy, n-butoxy, i-butoxy, sec-butoxy, tert-butoxycyclopropoxy, cyclohexyloxy, and the like.

The term “amine” or “amino” refers to a group of the formula —NRR′,where R and R′ can be, independently, hydrogen or an alkyl, alkenyl,alkynyl, aryl, aralkyl, cycloalkyl, halogenated alkyl, orheterocycloalkyl group. For example, an “alkylamino” or “alkylatedamino” refers to —NRR′, wherein at least one of R or R′ is an alkyl.

The term “aminoalkyl” refers to alkyl groups as defined above where atleast one hydrogen atom is replaced with an amino group (e.g.,—CH₂—NH₂).

“Aminocarbonyl” alone or in combination, means an amino substitutedcarbonyl (carbamoyl) radical, wherein the amino radical may optionallybe mono- or di-substituted, such as with alkyl, aryl, aralkyl,cycloalkyl, cycloalkylalkyl, alkanoyl, alkoxycarbonyl, aralkoxycarbonyland the like.

The term “amide” or “amido” is represented by the formula —C(O)NRR′,where R and R′ independently can be a hydrogen, alkyl, alkenyl, alkynyl,acyl, aryl, aralkyl, cycloalkyl, halogenated alkyl, or heterocycloalkylgroup described above. A suitable amido group is acetamido.

An “animal” refers to living multi-cellular vertebrate organisms, acategory that includes, for example, mammals and birds. The term mammalincludes both human and non-human mammals.

Similarly, the term “subject” includes both human and non-humansubjects, including birds and non-human mammals, such as non-humanprimates, companion animals (such as dogs and cats), livestock (such aspigs, sheep, cows), as well as non-domesticated animals, such as the bigcats. The term subject applies regardless of the stage in the organism'slife-cycle. Thus, the term subject applies to an organism in utero or inovo, depending on the organism (that is, whether the organism is amammal or a bird, such as a domesticated or wild fowl).

An “analog” is a molecule that differs in chemical structure from aparent compound, for example a homolog (differing by an increment in thechemical structure or mass, such as a difference in the length of analkyl chain or the inclusion of one of more isotopes), a molecularfragment, a structure that differs by one or more functional groups, ora change in ionization. An analog is not necessarily synthesized fromthe parent compound. Structural analogs are often found usingquantitative structure activity relationships (QSAR), with techniquessuch as those disclosed in Remington (The Science and Practice ofPharmacology, 19th Edition (1995), chapter 28). A derivative is amolecule derived from the base structure.

The term “aralkyl” refers to a group in which an aryl group issubstituted for a hydrogen atom of an alkyl group. An example of anaralkyl group is a benzyl group.

The term “aryl” refers to any group derived from an aromatic groupincluding, but not limited to, a benzene ring or to an optionallysubstituted benzene ring system fused to one or more optionallysubstituted benzene rings. A “heteroaryl group” is defined as an arylgroup that has at least one heteroatom incorporated within the ring ofthe aryl group. Examples of heteroatoms include, but are not limited to,nitrogen, oxygen, sulfur, and phosphorous. The aryl group can besubstituted with one or more groups including, but not limited to,alkyl, alkynyl, alkenyl, aryl, halide, nitro, amino, ester, ketone,aldehyde, hydroxy, carboxylic acid, or alkoxy, or the aryl group can beunsubstituted. Examples of an aryl group include, but are not limitedto, phenyl, 2-naphthyl, 1-naphthyl, 1-anthracenyl, and the like.

The term “arylene” refers to a divalent or higher valency benzene ringgroup or a divalent or higher valency benzene ring system fused to onemore optionally substituted benzene rings. The arylene group can besubstituted with one or more groups including, but not limited to,alkyl, alkynyl, alkenyl, aryl, halide, nitro, amino, ester, ketone,aldehyde, hydroxy, carboxylic acid, or alkoxy, or the arylene group canbe unsubstituted. Examples of an arylene group include, but are notlimited to, phenylene (e.g., benzene-1,4-diyl), naphthalene-1,8-diyl,benzenetriyl, benzenetetrayl, and the like.

“Carbonyl” refers to a radical of the formula —C(O)—.Carbonyl-containing groups include any substituent containing acarbon-oxygen double bond (C═O), including acyl groups, amides, carboxygroups, esters, ureas, carbamates, carbonates and ketones and aldehydes,such as substituents based on —COR or —RCHO where R is an aliphatic,heteroaliphatic, alkyl, heteroalkyl, hydroxyl, or a secondary, tertiary,or quaternary amine.

A carbonylamino group may be —N(R)—C(O)—R (wherein each R isindependently a substitution group such as, for example, alkyl, alkenyl,alkynyl, acyl, aryl, aralkyl, cycloalkyl, halogenated alkyl, orheterocycloalkyl group, or H).

“Carboxyl” refers to a —COOH radical. The carboxyl group can form acarboxylic acid. “Substituted carboxyl” refers to —COOR where R isalkyl, alkenyl, alkynyl, aryl, aralkyl, cycloalkyl, halogenated alkyl,or heterocycloalkyl group. For example, a substituted carboxyl groupcould be a carboxylic acid ester or a salt thereof (e.g., a carboxylate)

The term “cycloalkyl” refers to a non-aromatic carbon-based ringcomposed of at least three carbon atoms. Examples of cycloalkyl groupsinclude, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl,cyclohexyl, and the like. The term “heterocycloalkyl group” is acycloalkyl group as defined above where at least one of the carbon atomsof the ring is substituted with a heteroatom such as, but not limitedto, nitrogen, oxygen, sulfur, or phosphorous.

“Derivative”: In chemistry, a derivative is a compound that is derivedfrom a similar compound or a compound that can be imagined to arise fromanother compound, for example, if one atom is replaced with another atomor group of atoms. The latter definition is common in organic chemistry.In biochemistry, derivative refers to compounds that at leasttheoretically can be formed from the precursor compound.

“Drug-resistant” or “multidrug-resistant” refers to a cancer that isresistant to treatment by at least one therapeutic agent historicallyadministered to treat that cancer. These recurrent cancers often occurafter surgery, primary chemotherapy treatment, radiotherapy, orimmunotherapy. In certain embodiments, the cancer is achemotherapeutic-resistant carcinoma.

The terms “halogenated alkyl” or “haloalkyl group” refer to an alkylgroup as defined above with one or more hydrogen atoms present on thesegroups substituted with a halogen (F, Cl, Br, I).

The term “hydroxyl” is represented by the formula —OH.

The term “hydroxyalkyl” refers to an alkyl group that has at least onehydrogen atom substituted with a hydroxyl group. The term “alkoxyalkylgroup” is defined as an alkyl group that has at least one hydrogen atomsubstituted with an alkoxy group described above.

“Inhibiting” refers to inhibiting the full development of a disease orcondition. “Inhibiting” also refers to any quantitative or qualitativereduction in biological activity or binding, relative to a control.

The term “neoplasm” refers to an abnormal cellular proliferation, whichincludes benign and malignant tumors, as well as other proliferativedisorders.

“Optional” or “optionally” means that the subsequently described eventor circumstance can but need not occur, and that the descriptionincludes instances where said event or circumstance occurs and instanceswhere it does not.

The terms “pharmaceutically acceptable salt or ester” refers to salts oresters prepared by conventional means that include salts, e.g., ofinorganic and organic acids, including but not limited to hydrochloricacid, hydrobromic acid, sulfuric acid, phosphoric acid, methanesulfonicacid, ethanesulfonic acid, malic acid, acetic acid, oxalic acid,tartaric acid, citric acid, lactic acid, fumaric acid, succinic acid,maleic acid, salicylic acid, benzoic acid, phenylacetic acid, mandelicacid and the like. “Pharmaceutically acceptable salts” of the presentlydisclosed compounds also include those formed from cations such assodium, potassium, aluminum, calcium, lithium, magnesium, zinc, and frombases such as ammonia, ethylenediamine, N-methyl-glutamine, lysine,arginine, ornithine, choline, N,N′-dibenzylethylenediamine,chloroprocaine, diethanolamine, procaine, N-benzylphenethylamine,diethylamine, piperazine, tris(hydroxymethyl)aminomethane, andtetramethylammonium hydroxide. These salts may be prepared by standardprocedures, for example by reacting the free acid with a suitableorganic or inorganic base. Any chemical compound recited in thisspecification may alternatively be administered as a pharmaceuticallyacceptable salt thereof. “Pharmaceutically acceptable salts” are alsoinclusive of the free acid, base, and zwitterionic forms. Descriptionsof suitable pharmaceutically acceptable salts can be found in Handbookof Pharmaceutical Salts, Properties, Selection and Use, Wiley VCH(2002). When compounds disclosed herein include an acidic function suchas a carboxy group, then suitable pharmaceutically acceptable cationpairs for the carboxy group are well known to those skilled in the artand include alkaline, alkaline earth, ammonium, quaternary ammoniumcations and the like. Such salts are known to those of skill in the art.

For additional examples of “pharmacologically acceptable salts,” seeBerge et al., J. Pharm. Sci. 66:1 (1977).

“Pharmaceutically acceptable esters” includes those derived fromcompounds described herein that are modified to include a hydroxy or acarboxyl group. An in vivo hydrolysable ester is an ester, which ishydrolysed in the human or animal body to produce the parent acid oralcohol. Suitable pharmaceutically acceptable esters that include acarboxyl group include C₁₋₆ alkoxymethyl esters for examplemethoxy-methyl, C₁₋₆ alkanoyloxymethyl esters for examplepivaloyloxymethyl, phthalidyl esters, C₃₋₈ cycloalkoxycarbonyloxy, C₁₋₆alkyl esters for example 1-cyclohexylcarbonyl-oxyethyl;1,3-dioxolen-2-onylmethyl esters for example5-methyl-1,3-dioxolen-2-onylmethyl; and C₁₋₆ alkoxycarbonyloxyethylesters for example 1-methoxycarbonyl-oxyethyl which may be formed at anycarboxy group in the compounds.

An in vivo hydrolysable ester containing a hydroxy group includesinorganic esters such as phosphate esters and α-acyloxyalkyl ethers andrelated compounds which as a result of the in vivo hydrolysis of theester breakdown to give the parent hydroxy group. Examples ofα-acyloxyalkyl ethers include acetoxy-methoxy and2,2-dimethylpropionyloxy-methoxy. A selection of in vivo hydrolysableester forming groups for hydroxy include alkanoyl, benzoyl, phenylacetyland substituted benzoyl and phenylacetyl, alkoxycarbonyl (to give alkylcarbonate esters), dialkylcarbamoyl andN-(dialkylaminoethyl)-N-alkylcarbamoyl (to give carbamates),dialkylaminoacetyl and carboxyacetyl. Examples of substituents onbenzoyl include morpholino and piperazino linked from a ring nitrogenatom via a methylene group to the 3- or 4-position of the benzoyl ring.

For therapeutic use, salts of the compounds are those wherein thecounter-ion is pharmaceutically acceptable. However, salts of acids andbases which are non-pharmaceutically acceptable may also find use, forexample, in the preparation or purification of a pharmaceuticallyacceptable compound.

The pharmaceutically acceptable acid and base addition salts asmentioned hereinabove are meant to comprise the therapeutically activenon-toxic acid and base addition salt forms which the compounds are ableto form. The pharmaceutically acceptable acid addition salts canconveniently be obtained by treating the base form with such appropriateacid. Appropriate acids comprise, for example, inorganic acids such ashydrohalic acids, e.g. hydrochloric or hydrobromic acid, sulfuric,nitric, phosphoric and the like acids; or organic acids such as, forexample, acetic, propanoic, hydroxyacetic, lactic, pyruvic, oxalic (i.e.ethanedioic), malonic, succinic (i.e. butanedioic acid), maleic,fumaric, malic (i.e. hydroxybutanedioic acid), tartaric, citric,methanesulfonic, ethanesulfonic, benzenesulfonic, p-toluenesulfonic,cyclamic, salicylic, p-aminosalicylic, pamoic and the like acids.Conversely said salt forms can be converted by treatment with anappropriate base into the free base form.

The compounds containing an acidic proton may also be converted intotheir non-toxic metal or amine addition salt forms by treatment withappropriate organic and inorganic bases. Appropriate base salt formscomprise, for example, the ammonium salts, the alkali and earth alkalinemetal salts, e.g. the lithium, sodium, potassium, magnesium, calciumsalts and the like, salts with organic bases, e.g. the benzathine,N-methyl-D-glucamine, hydrabamine salts, and salts with amino acids suchas, for example, arginine, lysine and the like.

The term “addition salt” as used hereinabove also comprises the solvateswhich the compounds described herein are able to form. Such solvates arefor example hydrates, alcoholates and the like.

The term “quaternary amine” as used hereinbefore defines the quaternaryammonium salts which the compounds are able to form by reaction betweena basic nitrogen of a compound and an appropriate quatemizing agent,such as, for example, an optionally substituted alkylhalide, arylhalideor arylalkylhalide, e.g. methyliodide or benzyliodide. Other reactantswith good leaving groups may also be used, such as alkyltrifluoromethanesulfonates, alkyl methanesulfonates, and alkylp-toluenesulfonates. A quaternary amine has a positively chargednitrogen. Pharmaceutically acceptable counterions include chloro, bromo,iodo, trifluoroacetate and acetate. The counterion of choice can beintroduced using ion exchange resins.

It will be appreciated that the compounds described herein may havemetal binding, chelating, complex forming properties and therefore mayexist as metal complexes or metal chelates.

Some of the compounds described herein may also exist in theirtautomeric form.

“Preventing” a disease or condition refers to prophylactic administeringa composition to a subject who does not exhibit signs of a disease orexhibits only early signs for the purpose of decreasing the risk ofdeveloping a pathology or condition, or diminishing the severity of apathology or condition.

Prodrugs of the disclosed compounds also are contemplated herein. Aprodrug is an active or inactive compound that is modified chemicallythrough in vivo physiological action, such as hydrolysis, metabolism andthe like, into an active compound following administration of theprodrug to a subject. The term “prodrug” as used throughout this textmeans the pharmacologically acceptable derivatives such as esters,amides and phosphates, such that the resulting in vivo biotransformationproduct of the derivative is the active drug as defined in the compoundsdescribed herein. Prodrugs preferably have excellent aqueous solubility,increased bioavailability and are readily metabolized into the activeinhibitors in vivo. Prodrugs of a compounds described herein may beprepared by modifying functional groups present in the compound in sucha way that the modifications are cleaved, either by routine manipulationor in vivo, to the parent compound. The suitability and techniquesinvolved in making and using prodrugs are well known by those skilled inthe art. F or a general discussion of prodrugs involving esters seeSvensson and Tunek, Drug Metabolism Reviews 165 (1988) and Bundgaard,Design of Prodrugs, Elsevier (1985).

The term “prodrug” also is intended to include any covalently bondedcarriers that release an active parent drug of the present invention invivo when the prodrug is administered to a subject. Since prodrugs oftenhave enhanced properties relative to the active agent pharmaceutical,such as, solubility and bioavailability, the compounds disclosed hereincan be delivered in prodrug form. Thus, also contemplated are prodrugsof the presently disclosed compounds, methods of delivering prodrugs andcompositions containing such prodrugs. Prodrugs of the disclosedcompounds typically are prepared by modifying one or more functionalgroups present in the compound in such a way that the modifications arecleaved, either in routine manipulation or in vivo, to yield the parentcompound. Prodrugs include compounds having a phosphonate and/or aminogroup functionalized with any group that is cleaved in vivo to yield thecorresponding amino and/or phosphonate group, respectively. Examples ofprodrugs include, without limitation, compounds having an acylated aminogroup and/or a phosphonate ester or phosphonate amide group. Inparticular examples, a prodrug is a lower alkyl phosphonate ester, suchas an isopropyl phosphonate ester.

Protected derivatives of the disclosed compounds also are contemplated.A variety of suitable protecting groups for use with the disclosedcompounds are disclosed in Greene and Wuts, Protective Groups in OrganicSynthesis; 3rd Ed.; John Wiley & Sons, New York, 1999.

In general, protecting groups are removed under conditions which willnot affect the remaining portion of the molecule. These methods are wellknown in the art and include acid hydrolysis, hydrogenolysis and thelike. One preferred method involves the removal of an ester, such ascleavage of a phosphonate ester using Lewis acidic conditions, such asin TMS-Br mediated ester cleavage to yield the free phosphonate. Asecond preferred method involves removal of a protecting group, such asremoval of a benzyl group by hydrogenolysis utilizing palladium oncarbon in a suitable solvent system such as an alcohol, acetic acid, andthe like or mixtures thereof. A t-butoxy-based group, including t-butoxycarbonyl protecting groups can be removed utilizing an inorganic ororganic acid, such as HCl or trifluoroacetic acid, in a suitable solventsystem, such as water, dioxane and/or methylene chloride. Anotherexemplary protecting group, suitable for protecting amino and hydroxyfunctions amino is trityl. Other conventional protecting groups areknown and suitable protecting groups can be selected by those of skillin the art in consultation with Greene and Wuts, Protective Groups inOrganic Synthesis; 3rd Ed.; John Wiley & Sons, New York, 1999. When anamine is deprotected, the resulting salt can readily be neutralized toyield the free amine. Similarly, when an acid moiety, such as aphosphonic acid moiety is unveiled, the compound may be isolated as theacid compound or as a salt thereof.

The term “subject” includes both human and veterinary subjects.

“Substituted” or “substitution” refers to replacement of a hydrogen atomof a molecule or an R-group with one or more additional R-groups. Unlessotherwise defined, the term “optionally-substituted” or “optionalsubstituent” as used herein refers to a group which may or may not befurther substituted with 1, 2, 3, 4 or more groups, preferably 1, 2 or3, more preferably 1 or 2 groups. The substituents may be selected, forexample, from C₁₋₆alkyl, C₂₋₆alkenyl, C₂₋₆alkynyl, C₃₋₈cycloalkyl,hydroxyl, oxo, C₁₋₆alkoxy, aryloxy, C₁₋₆alkoxyaryl, halo, C₁₋₆alkylhalo(such as CF₃ and CHF₂), C₁₋₆alkoxyhalo (such as OCF₃ and OCHF₂),carboxyl, esters, cyano, nitro, amino, substituted amino, disubstitutedamino, acyl, ketones, amides, aminoacyl, substituted amides,disubstituted amides, thiol, alkylthio, thioxo, sulfates, sulfonates,sulfinyl, substituted sulfinyl, sulfonyl, substituted sulfonyl,sulfonylamides, substituted sulfonamides, disubstituted sulfonamides,aryl, arC₁₋₆alkyl, heterocyclyl and heteroaryl wherein each alkyl,alkenyl, alkynyl, cycloalkyl, aryl and heterocyclyl and groupscontaining them may be further optionally substituted. Optionalsubstituents in the case N-heterocycles may also include but are notlimited to C₁₋₆alkyl i.e. N—C₁₋₃alkyl, more preferably methylparticularly N-methyl.

A “therapeutically effective amount” refers to a quantity of a specifiedagent sufficient to achieve a desired effect in a subject being treatedwith that agent. Ideally, a therapeutically effective amount of an agentis an amount sufficient to inhibit or treat the disease or conditionwithout causing a substantial cytotoxic effect in the subject. Thetherapeutically effective amount of an agent will be dependent on thesubject being treated, the severity of the affliction, and the manner ofadministration of the therapeutic composition.

“Treatment” refers to a therapeutic intervention that ameliorates a signor symptom of a disease or pathological condition after it has begun todevelop. As used herein, the term “ameliorating,” with reference to adisease or pathological condition, refers to any observable beneficialeffect of the treatment. The beneficial effect can be evidenced, forexample, by a delayed onset of clinical symptoms of the disease in asusceptible subject, a reduction in severity of some or all clinicalsymptoms of the disease, a slower progression of the disease, animprovement in the overall health or well-being of the subject, or byother parameters well known in the art that are specific to theparticular disease. The phrase “treating a disease” is inclusive ofinhibiting the full development of a disease or condition, for example,in a subject who is at risk for a disease, or who has a disease, such ascancer, particularly a metastatic cancer.

Inhibitors

Unless context clearly indicates otherwise, all compounds describedherein may be provided as a pharmaceutically acceptable salt thereof. Insome embodiments, the inhibitor is not a salt. In some embodiments, theinhibitor is a salt. In certain embodiments, the inhibitors may be lowmolecular weight compounds (“LMWCs”, having a molecular weight of lessthan about, for example and not by way of limitation, 600 daltons).

Particular examples of the presently disclosed compounds may include oneor more asymmetric centers; thus these compounds can exist in differentstereoisomeric forms. Accordingly, compounds and compositions may beprovided as individual pure enantiomers or as stereoisomeric mixtures,including racemic mixtures. In certain embodiments the compoundsdisclosed herein are synthesized in or are purified to be insubstantially enantiopure form, such as in a 90% enantiomeric excess, a95% enantiomeric excess, a 97% enantiomeric excess or even in greaterthan a 99% enantiomeric excess, such as in enantiopure form.

The presently disclosed compounds can have at least one asymmetriccenter or geometric center, cis-trans center (C═C, C═N). All chiral,diasteromeric, racemic, meso, rotational and geometric isomers of thestructures are intended unless otherwise specified. The compounds can beisolated as a single isomer or as mixture of isomers. All tautomers ofthe compounds are also considered part of the disclosure. The presentlydisclosed compounds also includes all isotopes of atoms present in thecompounds, which can include, but are not limited to, deuterium,tritium, ¹⁸F, etc

Disclosed herein are compounds that are p62-ZZ inhibitors having aformula I of:

-   -   wherein Ar is an arylene or heteroarylene group;        R¹ has a structure of:

-   -   wherein W is an alkanediyl, alkenediyl, a carbonyl, or a        combination thereof;    -   X is —NR⁵—, wherein R⁵ is H or an alkyl, or —O—; and    -   Y is optionally-substituted cycloalkyl, optionally-substituted        cycloalkyl-substituted alkyl, optionally-substituted        heterocycloalkyl, or optionally-substituted        heterocycloalkyl-substituted alkyl;        each R² is the same or different and has a structure of:

-   -   wherein Z is —NR⁶—, wherein R⁶ is H or an alkyl, —O—, —S—, or        —CH₂—;    -   Z¹ is (—CH₂—)_(m) wherein m is 0 to 5, or an alkenediyl having 2        to 6 carbon atoms;    -   Cy is a 3-8-membered cycloalkyl, heterocycloalkyl, aryl or        heteroaryl ring; and    -   each R⁴ is the same or different and is selected from hydroxy,        halogen, substituted or unsubstituted alkoxy, substituted or        unsubstituted alkyl, or amino; and c is 0 to 5; and    -   each R³ is the same or different and is selected from hydroxy,        halogen, substituted or unsubstituted alkoxy, substituted or        unsubstituted alkyl, amino, substituted or unsubstituted        cycloalkyl, substituted or unsubstituted heterocycloalkyl,        substituted or unsubstituted aryl, or a nitro, wherein a is 2 to        5, and b is 0 to 3.

Ar, for example, may be a benzenetriyl (e.g., a benzene-1,3,4-triyl) ora benzenetetrayl.

W, for example, may be a lower alkanediyl, or a lower alkenediyl, or alower alkanediyl that also includes a carbonyl, or a lower alkenediylthat also includes a carbonyl. X, for example, may be —NR⁵—, wherein R⁵is a lower alkyl, which may be optionally substituted. Y, for example,may be a cycloalkyl-substituted alkyl wherein the alkyl group includes 1to 4 carbon atoms. In certain embodiments, R¹ is —CH₂—X—(CH₂)_(m)—R¹¹,wherein X is NH or O, m is 0 to 6; and R¹¹ is optionally-substitutedcycloalkyl (particularly optionally-substituted cyclohexyl). In certainembodiments, R¹¹ is unsubstituted cyclohexyl.

In certain embodiments, Y is:

In certain embodiments, R² has a structure of:

Z, for example, may be —NR⁶—, wherein R⁶ is a lower alkyl, which may beoptionally substituted, or Z may be —O—. R⁴, for example, may be —F,—Cl, —OCH₃, —OH, —CH₃ or —NH₂. In certain embodiments, c is 1 or 2. Incertain embodiments, R⁴ is 4-fluoro; 2,4-difluoro; or 4-methyl.

R³, for example, may be —F, —Cl, —OCH₃, —OH, or —NH₂. In certainembodiments, R³ is a lower alkyl, a lower alkoxy, C₁-C₄ alkylamino (theC₁-C₄ alkyl moiety of the alkylamino may be straight-chained orbranched), di (C₁-C₄ alkyl)amino, C₃-C₇ cycloalkyl (particularly C₃-C₅cycloalkyl, especially cyclopropyl), a hydroxyl-substituted C₃-C₇cycloalkyl, a 3 to 6-membered heterocycloalkyl which contains at leastone heteroatom selected from N, O and S; an alkyl-substituted 3 to6-membered heterocycloalkyl, a hydroxyl-substituted 3 to 6-memberedheterocycloalkyl, or aryl such as phenyl, naphthyl or anthracenyl, or anitro group.

In certain embodiments, a is 2, 3, 4 or 5. In preferred embodiments, ais 2. In certain embodiments, a first R² group is in a meta position onthe Ar ring relative to the R¹ group, and a second R² group is in a paraposition on the Ar ring relative to the R¹ group. In certainembodiments, each R² group has the same structure.

In certain embodiments, b is 0, 1, 2 or 3.

In certain embodiments, c is 0, 1, 2, 3, 4 or 5.

In more particular embodiments, the p62-ZZ inhibitor is selected from:

-   -   wherein each of R⁷, R⁸, and R⁹ are the same or different and are        selected from —F, —Cl, —OCH₃, —OH, —CH₃, or —NH₂; d is 0 to 3; e        is 0 to 5; and f is 0 to 5.

The compounds disclosed herein may be synthesized as shown below.

General Synthesis Route:

Synthesis of Compound XIELP1-106:

First, 3,4-dihydroxybenzaldehyde (1.38 g, 10.0 mmol) was diluted withdry dimethylformamide (DMF, 40 mL). 1-(Bromomethyl)-4-fluorobenzene(3.96 g, 21.0 mmol) was added slowly, followed by anhydrous K₂CO₃ (5.52g, 40.0 mmol). The mixture was stirred at room temperature for 6 hours.The mixture was partitioned between H₂O and ether (120 mL each). Theorganic layer was separated and the water layer was extracted with ether(3×50 mL). The pooled organic layers were washed with H₂O (2×50 mL) andsaturated aqueous NaCl (50 mL). The pale, straw-colored extracts weredried over anhydrous sodium sulfate and concentrated to yield a whitecream-colored solid 3,4-bis((4-fluorobenzyl)oxy)benzaldehyde (4.31 g,82%) after washing with hexanes (75 mL) and drying.

Subsequently, 3,4-bis((4-fluorobenzyl)oxy)benzaldehyde (354 mg, 1 mmol)was dissolved in dry ethanol, and cyclohexylmethanamine (0.13 mL, 1mmol) was added. The reaction mixture was stirred for 12 hours at 60° C.The reaction solution was cooled down to room temperature. NaBH₄ (57 mg,1.5 mmol) was added slowly in small portions, and the resulting solutionwas stirred for another 12 hours. The solvent was evaporated in vacuum,and the residue was dissolved in water and extracted with ethyl acetate.The organic layers were combined and dried with Na₂SO₄, filtered, andevaporated in a vacuum. The residue was purified by a flash column togenerate the desired productN-(3,4-bis((4-fluorobenzyl)oxy)benzyl)-1-cyclohexylmethanamine (260 mg,57%).

Finally, 2-((3,4-bis(benzyloxy)benzyl)amino)ethan-1-ol (260 mg, 0.58mmol) was dissolved in 25 mL of absolute methanol and pumped with HClgas for 1 hour. The mixture was stirred for another 2 hours andevaporated to about 1 mL. Hexane was then added to get a solid compound,which was filtered and dried to give the final compound (210 mg, 74%).¹H NMR (CDCl₃): 7.52-7.33 (m, 10H), 7.01-6.84 (m, 3H), 5.20 (s, 2H),5.17 (s, 2H), 3.71 (s, 2H), 3.64 (t, J=4.8, 2H), 2.93 (s, 2H), 2.72 (t,J=4.8, 2H).

Treatment and Pharmaceutical Compositions

The compounds disclosed herein may be useful in treating p62-mediateddiseases. Illustrative p62-mediated diseases include multiple myeloma,autophagy-related diseases, metabolic syndrome, Alzheimer's disease andother neurodegenerative diseases, and infectious diseases such as thosecaused by Staphylococcus aureus, Enteroccocus, or Salmonella enterica.

The compounds disclosed herein may be useful in treating cancer such asrenal cancer, lung cancer, thyroid cancer, prostate cancer, multiplemyeloma and breast cancer.

In certain embodiments, the compounds disclosed herein inhibit multiplemyeloma cell growth. In particular, the compounds may be used fortreating multiple myeloma characterized by a cell type selected from oneor more OPM-2 cells, OPM-2-like cells, MM-IS cells, MM-IS-like cells,MM.1R cells, MM-1R-like cells, KMS-18 cells, KMS-18-like cells, S6B45cells, S6B45-like cells, MR20 cells, MR20-like cells, ARD cells and/orARD-like cells. Depending on the type of tumor and the development stageof the disease, anticancer effects of the methods of treatment include,but are not limited to, inhibition of tumor growth, tumor growth delay,regression of tumor, shrinkage of tumor, increased time to regrowth oftumor on cessation of treatment, slowing of disease progression, andprevention of metastasis. It is expected that when a method of treatmentis administered to a subject in need of such treatment, said method oftreatment will produce an effect, as measured by, for example, theextent of the anticancer effect, the response rate, the time to diseaseprogression, or the survival rate. In particular, the methods oftreatment are suited for human patients, especially those who arerelapsing or refractory to previous chemotherapy, although first linetherapy is also envisaged. For example, the compounds disclosed hereinmay be used for treating cancer, particularly drug-resistant multiplemyeloma that is resistant to one or more of dexamethasone, alkylatingagents (e.g., melphalan, cyclophosamide), anthracyclines (e.g.,doxorubicin), thalidomide, lenalidomide, CC-4047, bortezomib, andmultitargeted kinase inhibitors. The compound disclosed herein may alsobe co-administered with any of the above-referenced agents. Thecompounds disclosed herein may block autophagy. Blocking autophagy canenhance the efficacy of a DNA damaging agent for treatment of cancer(Suppression of autophagy by FIP200 deletion impairs DNA damage repairand increases cell death upon treatments with anti-cancer agents. Bae etal., Jl. Mol Cancer Res. 2011 Aug. 1) and possibly other agents.

In certain embodiments, the compounds disclosed herein modulate p62activity in stromal cells which can decrease tumor growth and bonedestruction.

In particular embodiments, the compounds disclosed herein may be usedfor inhibiting osteoclastogenesis and/or reducing osteoclast activation.Osteoclast is the primary bone-resorbing cell in both normal andpathologic states. Increased osteoclastic bone resorption can resultfrom both increased osteoclast formation and activation of preformedosteoclasts to resorb bone. In patients with bone metastases, osteolyticbone destruction can result in severe bone pain, pathologic fractures,hypercalcemia, and nerve compression syndromes. Several tumors show ahigh predilection for bone, including renal cancer, lung cancer, thyroidcancer, prostate cancer, multiple myeloma and breast cancer, see e.g.Roodman, Journal of Clinical Oncology, vol. 19, 2001, p. 3562.Osteoclast formation and activation may also contribute to osteolyticdisease and bone loss in individuals suffering from osteoporosis, suchas post-menopausal osteoporosis, Paget's disease, rheumatoid arthritisand head and neck squamous cell carcinoma, see e.g. U.S. Pat. No.7,462,646.

The compounds disclosed may be co-administered with another therapeuticagent, especially another anti-cancer agent. In certain embodiments,there is provided a pharmaceutical composition that includes at leastp62-ZZ inhibitor and at least one other anti-cancer agent. Illustrativechemotherapeutic agents include an EGF-receptor antagonist, arsenicsulfide, adriamycin, cisplatin, carboplatin, cimetidine, caminomycin,mechlorethamine hydrochloride, pentamethylmelamine, thiotepa,teniposide, cyclophosphamide, chlorambucil, demethoxyhypocrellin A,melphalan, ifosfamide, trofosfamide, Treosulfan, podophyllotoxin orpodophyllotoxin derivatives, etoposide phosphate, teniposide, etoposide,leurosidine, leurosine, vindesine, 9-aminocamptothecin,camptoirinotecan, crisnatol, megestrol, methopterin, mitomycin C,ecteinascidin 743, busulfan, carmustine, lomustine, lovastatin,1-methyl-4-phenylpyridinium ion, semustine, staurosporine, streptozocin,phthalocyanine, dacarbazine, aminopterin, methotrexate, trimetrexate,thioguanine, mercaptopurine, fludarabine, pentastatin, cladribin,cytarabine, porfiromycin, 5-fluorouracil, 6-mercaptopurine, doxorubicinhydrochloride, leucovorin, mycophenolic acid, daunorubicin,deferoxamine, floxuridine, doxifluridine, raltitrexed, idarubicin,epirubican, pirarubican, zorubicin, mitoxantrone, bleomycin sulfate,actinomycin D, safracins, saframycins, quinocarcins, discodermolides,vincristine, vinblastine, vinorelbine tartrate, vertoporfin, paclitaxel,tamoxifen, raloxifene, tiazofuran, thioguanine, ribavirin, EICAR,estramustine, estramustine phosphate sodium, flutamide, bicalutamide,buserelin, leuprolide, pteridines, enediynes, levamisole, aflacon,interferon, interleukins, aldesleukin, filgrastim, sargramostim,rituximab, BCG, tretinoin, betamethasone, gemcitabine hydrochloride,verapamil, VP-16, altretamine, thapsigargin, oxaliplatin, iproplatin,tetraplatin, lobaplatin, DCP, PLD-147, JM118, JM216, JM335, satraplatin,docetaxel, deoxygenated paclitaxel, TL-139, 5′-nor-anhydrovinblastine,camptothecin, irinotecan, topotecan, BAY 38-3441, 9-nitrocamptothecin,exatecan, lurtotecan, gimatecan, homocamptothecins diflomotecan and9-aminocamptothecin, SN-38, ST 1481, karanitecin, indolocarbazoles,protoberberines, intoplicines, idenoisoquinolones, benzo-phenazines andNB-506.

The compounds disclosed herein may also be administered to a subject inneed thereof to reduce food intake and body weight, reverse insulin andleptin resistance, reverse hepatic steatosis (fatty liver) and improvedyslipidemia. They may be used for treating obesity, diabetes (e.g.,type 2 diabetes), and non-alcoholic and alcoholic fatty liver disease(NAFLD/AFLD), the latter being a risk factor for insulin resistance,cirrhosis and liver cancer, dyslipidemias that predispose toarteriosclerotic heart disease, diabetic nephropathy, gout, andfibrosis.

The diabetes disorder may be Type 1 diabetes, Type 2 diabetes,inadequate glucose tolerance, and/or insulin resistance.

The compounds disclosed herein may also be administered to a subject inneed thereof for treating a neurodegenerative disease or disorder suchas, for example, Alzheimer's disease, ataxia telangiectasia, Parkinson'sdisease, amyotrophic lateral sclerosis, and Huntington's disease.

The compounds identified herein may be included in a pharmaceuticalcomposition that includes at least one pharmaceutically acceptableadditive such as a carrier, thickener, diluent, buffer, preservative,surface active agent and the like in addition to the agent.Pharmaceutical compositions can also include one or more additionalactive ingredients such as antimicrobial agents, anti-inflammatoryagents, anesthetics, and the like. The pharmaceutically acceptablecarriers useful for these formulations are conventional. Remington'sPharmaceutical Sciences, by E. W. Martin, Mack Publishing Co., Easton,Pa., 19th Edition (1995), describes compositions and formulationssuitable for pharmaceutical delivery of the compounds herein disclosed.

In general, the nature of the carrier will depend on the particular modeof administration being employed. For instance, parenteral formulationsusually contain injectable fluids that include pharmaceutically andphysiologically acceptable fluids such as water, physiological saline,balanced salt solutions, aqueous dextrose, glycerol or the like as avehicle. For solid compositions (for example, powder, pill, tablet, orcapsule forms), conventional non-toxic solid carriers can include, forexample, pharmaceutical grades of mannitol, lactose, starch, ormagnesium stearate. In addition to biologically-neutral carriers,pharmaceutical compositions to be administered can contain minor amountsof non-toxic auxiliary substances, such as wetting or emulsifyingagents, preservatives, and pH buffering agents and the like, for examplesodium acetate or sorbitan monolaurate.

The compounds disclosed herein can be administered to subjects by avariety of mucosal administration modes, including by oral, rectal,intranasal, intrapulmonary, or transdermal delivery, or by topicaldelivery to other surfaces. Optionally, the compounds can beadministered by non-mucosal routes, including by intramuscular,subcutaneous, intravenous, intra-arterial, intra-articular,intraperitoneal, intrathecal, intracerebroventricular, or parenteralroutes. In other alternative embodiments, the compounds can beadministered ex vivo by direct exposure to cells, tissues or organsoriginating from a subject.

To formulate the pharmaceutical compositions, the compounds can becombined with various pharmaceutically acceptable additives, as well asa base or vehicle for dispersion of the compound. Desired additivesinclude, but are not limited to, pH control agents, such as arginine,sodium hydroxide, glycine, hydrochloric acid, citric acid, and the like.In addition, local anesthetics (for example, benzyl alcohol),isotonizing agents (for example, sodium chloride, mannitol, sorbitol),adsorption inhibitors (for example, Tween 80 or Miglyol 812), solubilityenhancing agents (for example, cyclodextrins and derivatives thereof),stabilizers (for example, serum albumin), and reducing agents (forexample, glutathione) can be included.

Adjuvants, such as aluminum hydroxide (for example, Amphogel, WyethLaboratories, Madison, N.J.), Freund's adjuvant, MPL™ (3-O-deacylatedmonophosphoryl lipid A; Corixa, Hamilton, Ind.) and IL-12 (GeneticsInstitute, Cambridge, Mass.), among many other suitable adjuvants wellknown in the art, can be included in the compositions. When thecomposition is a liquid, the tonicity of the formulation, as measuredwith reference to the tonicity of 0.9% (w/v) physiological salinesolution taken as unity, is typically adjusted to a value at which nosubstantial, irreversible tissue damage will be induced at the site ofadministration. Generally, the tonicity of the solution is adjusted to avalue of about 0.3 to about 3.0, such as about 0.5 to about 2.0, orabout 0.8 to about 1.7.

The compounds can be dispersed in a base or vehicle, which can include ahydrophilic compound having a capacity to disperse the compound, and anydesired additives. The base can be selected from a wide range ofsuitable compounds, including but not limited to, copolymers ofpolycarboxylic acids or salts thereof, carboxylic anhydrides (forexample, maleic anhydride) with other monomers (for example, methyl(meth)acrylate, acrylic acid and the like), hydrophilic vinyl polymers,such as polyvinyl acetate, polyvinyl alcohol, polyvinylpyrrolidone,cellulose derivatives, such as hydroxymethylcellulose,hydroxypropylcellulose and the like, and natural polymers, such aschitosan, collagen, sodium alginate, gelatin, hyaluronic acid, andnontoxic metal salts thereof. Often, a biodegradable polymer is selectedas a base or vehicle, for example, polylactic acid, poly(lacticacid-glycolic acid) copolymer, polyhydroxybutyric acid,poly(hydroxybutyric acid-glycolic acid) copolymer and mixtures thereof.Alternatively or additionally, synthetic fatty acid esters such aspolyglycerin fatty acid esters, sucrose fatty acid esters and the likecan be employed as vehicles. Hydrophilic polymers and other vehicles canbe used alone or in combination, and enhanced structural integrity canbe imparted to the vehicle by partial crystallization, ionic bonding,cross-linking and the like. The vehicle can be provided in a variety offorms, including fluid or viscous solutions, gels, pastes, powders,microspheres and films for direct application to a mucosal surface.

The compounds can be combined with the base or vehicle according to avariety of methods, and release of the compounds can be by diffusion,disintegration of the vehicle, or associated formation of waterchannels. In some circumstances, the compound is dispersed inmicrocapsules (microspheres) or nanocapsules (nanospheres) prepared froma suitable polymer, for example, isobutyl 2-cyanoacrylate (see, forexample, Michael et al., J. Pharmacy Pharmacol. 43:1-5, 1991), anddispersed in a biocompatible dispersing medium, which yields sustaineddelivery and biological activity over a protracted time.

The compositions of the disclosure can alternatively contain aspharmaceutically acceptable vehicles substances as required toapproximate physiological conditions, such as pH adjusting and bufferingagents, tonicity adjusting agents, wetting agents and the like, forexample, sodium acetate, sodium lactate, sodium chloride, potassiumchloride, calcium chloride, sorbitan monolaurate, and triethanolamineoleate. For solid compositions, conventional nontoxic pharmaceuticallyacceptable vehicles can be used which include, for example,pharmaceutical grades of mannitol, lactose, starch, magnesium stearate,sodium saccharin, talcum, cellulose, glucose, sucrose, magnesiumcarbonate, and the like.

Pharmaceutical compositions for administering the compounds can also beformulated as a solution, microemulsion, or other ordered structuresuitable for high concentration of active ingredients. The vehicle canbe a solvent or dispersion medium containing, for example, water,ethanol, polyol (for example, glycerol, propylene glycol, liquidpolyethylene glycol, and the like), and suitable mixtures thereof.Proper fluidity for solutions can be maintained, for example, by the useof a coating such as lecithin, by the maintenance of a desired particlesize in the case of dispersible formulations, and by the use ofsurfactants. In many cases, it will be desirable to include isotonicagents, for example, sugars, polyalcohols, such as mannitol andsorbitol, or sodium chloride in the composition. Prolonged absorption ofthe compound can be brought about by including in the composition anagent which delays absorption, for example, monostearate salts andgelatin.

In certain embodiments, the compounds can be administered in a timerelease formulation, for example in a composition which includes a slowrelease polymer. These compositions can be prepared with vehicles thatwill protect against rapid release, for example a controlled releasevehicle such as a polymer, microencapsulated delivery system orbioadhesive gel. Prolonged delivery in various compositions of thedisclosure can be brought about by including in the composition agentsthat delay absorption, for example, aluminum monostearate hydrogels andgelatin. When controlled release formulations are desired, controlledrelease binders suitable for use in accordance with the disclosureinclude any biocompatible controlled release material which is inert tothe active agent and which is capable of incorporating the compoundand/or other biologically active agent. Numerous such materials areknown in the art. Useful controlled-release binders are materials thatare metabolized slowly under physiological conditions following theirdelivery (for example, at a mucosal surface, or in the presence ofbodily fluids). Appropriate binders include, but are not limited to,biocompatible polymers and copolymers well known in the art for use insustained release formulations. Such biocompatible compounds arenon-toxic and inert to surrounding tissues, and do not triggersignificant adverse side effects, such as nasal irritation, immuneresponse, inflammation, or the like. They are metabolized into metabolicproducts that are also biocompatible and easily eliminated from thebody.

Exemplary polymeric materials for use in the present disclosure include,but are not limited to, polymeric matrices derived from copolymeric andhomopolymeric polyesters having hydrolyzable ester linkages. A number ofthese are known in the art to be biodegradable and to lead todegradation products having no or low toxicity. Exemplary polymersinclude polyglycolic acids and polylactic acids, poly(DL-lacticacid-co-glycolic acid), poly(D-lactic acid-co-glycolic acid), andpoly(L-lactic acid-co-glycolic acid). Other useful biodegradable orbioerodable polymers include, but are not limited to, such polymers aspoly(epsilon-caprolactone), poly(epsilon-aprolactone-CO-lactic acid),poly(epsilon.-aprolactone-CO-glycolic acid), poly(beta-hydroxy butyricacid), poly(alkyl-2-cyanoacrilate), hydrogels, such as poly(hydroxyethylmethacrylate), polyamides, poly(amino acids) (for example, L-leucine,glutamic acid, L-aspartic acid and the like), poly(ester urea),poly(2-hydroxyethyl DL-aspartamide), polyacetal polymers,polyorthoesters, polycarbonate, polymaleamides, polysaccharides, andcopolymers thereof. Many methods for preparing such formulations arewell known to those skilled in the art (see, for example, Sustained andControlled Release Drug Delivery Systems, J. R. Robinson, ed., MarcelDekker, Inc., New York, 1978). Other useful formulations includecontrolled-release microcapsules (U.S. Pat. Nos. 4,652,441 and4,917,893), lactic acid-glycolic acid copolymers useful in makingmicrocapsules and other formulations (U.S. Pat. Nos. 4,677,191 and4,728,721) and sustained-release compositions for water-soluble peptides(U.S. Pat. No. 4,675,189).

The pharmaceutical compositions of the disclosure typically are sterileand stable under conditions of manufacture, storage and use. Sterilesolutions can be prepared by incorporating the compound in the requiredamount in an appropriate solvent with one or a combination ofingredients enumerated herein, as required, followed by filteredsterilization. Generally, dispersions are prepared by incorporating thecompound and/or other biologically active agent into a sterile vehiclethat contains a basic dispersion medium and the required otheringredients from those enumerated herein. In the case of sterilepowders, methods of preparation include vacuum drying and freeze-dryingwhich yields a powder of the compound plus any additional desiredingredient from a previously sterile-filtered solution thereof. Theprevention of the action of microorganisms can be accomplished byvarious antibacterial and antifungal agents, for example, parabens,chlorobutanol, phenol, sorbic acid, thimerosal, and the like.

In accordance with the various treatment methods of the disclosure, thecompounds can be delivered to a subject in a manner consistent withconventional methodologies associated with management of the disorderfor which treatment or prevention is sought. In accordance with thedisclosure herein, a prophylactically or therapeutically effectiveamount of the compound is administered to a subject in need of suchtreatment for a time and under conditions sufficient to prevent,inhibit, and/or ameliorate a selected disease or condition or one ormore symptom(s) thereof.

The administration of the compounds can be for either prophylactic ortherapeutic purpose. When provided prophylactically, the compounds areprovided in advance of any symptom. The prophylactic administration ofthe compounds serves to prevent or ameliorate any subsequent diseaseprocess. When provided therapeutically, the compound is provided at (orshortly after) the onset of a symptom of disease or infection.

For prophylactic and therapeutic purposes, the compounds can beadministered to the subject by the oral route or in a single bolusdelivery, via continuous delivery (for example, continuous transdermal,mucosal or intravenous delivery) over an extended time period, or in arepeated administration protocol (for example, by an hourly, daily orweekly, repeated administration protocol). The therapeutically effectivedosage of the compound can be provided as repeated doses within aprolonged prophylaxis or treatment regimen that will yield clinicallysignificant results to alleviate one or more symptoms or detectableconditions associated with a targeted disease or condition as set forthherein. Determination of effective dosages in this context is typicallybased on animal model studies followed up by human clinical trials andis guided by administration protocols that significantly reduce theoccurrence or severity of targeted disease symptoms or conditions in thesubject. Suitable models in this regard include, for example, murine,rat, avian, porcine, feline, non-human primate, and other acceptedanimal model subjects known in the art. Alternatively, effective dosagescan be determined using in vitro models. Using such models, onlyordinary calculations and adjustments are required to determine anappropriate concentration and dose to administer a therapeuticallyeffective amount of the compound (for example, amounts that areeffective to elicit a desired immune response or alleviate one or moresymptoms of a targeted disease). In alternative embodiments, aneffective amount or effective dose of the agents may simply inhibit orenhance one or more selected biological activities correlated with adisease or condition, as set forth herein, for either therapeutic ordiagnostic purposes.

The actual dosage of the compounds will vary according to factors suchas the disease indication and particular status of the subject (forexample, the subject's age, size, fitness, extent of symptoms,susceptibility factors, and the like), time and route of administration,other drugs or treatments being administered concurrently, as well asthe specific pharmacology of the agent for eliciting the desiredactivity or biological response in the subject. Dosage regimens can beadjusted to provide an optimum prophylactic or therapeutic response. Atherapeutically effective amount is also one in which any toxic ordetrimental side effects of the compound are outweighed in clinicalterms by therapeutically beneficial effects. A non-limiting range for atherapeutically effective amount of a compound within the methods andformulations of the disclosure is about 0.01 mg/kg body weight to about20 mg/kg body weight, such as about 0.05 mg/kg to about 5 mg/kg bodyweight, or about 0.2 mg/kg to about 2 mg/kg body weight.

Dosage can be varied by the attending clinician to maintain a desiredconcentration at a target site (for example, the lungs or systemiccirculation). Higher or lower concentrations can be selected based onthe mode of delivery, for example, trans-epidermal, rectal, oral,pulmonary, or intranasal delivery versus intravenous or subcutaneousdelivery. Dosage can also be adjusted based on the release rate of theadministered formulation, for example, of an intrapulmonary spray versuspowder, sustained release oral versus injected particulate ortransdermal delivery formulations, and so forth.

EXAMPLES

It is known that the marrow microenvironment provides a criticalsupportive role in MM and enhances both tumor growth and bonedestruction through activation of multiple signaling pathways in stromalcells. Sequestosome 1 (p62) plays a key role in the formation ofsignaling complexes that result in NF-KB, p38 MAPK and PI3K activationin the marrow microenvironment of patients with MM. Deletion constructsof p62 were generated that lacked specific p62 domains: ΔSH2, ΔPB1, ΔZZ,Δp38, ΔTBS and ΔUBA to identify the domains, particular ZZ domain of p62responsible for increased MM cell growth and osteoclast (OCL) formationmediated by NF-KB and p38 MAPK signaling, as a means to developinhibitory peptides/molecules as potential therapeutic agents for MM(FIG. 1). These constructs were then transfected into a p62-knockoutstromal cell-line and it was found that the ZZ domain of p62 is requiredfor stromal cell support of MM cell growth, increased IL-6, VCAM-1expression and OCL formation. These results suggest that dominantnegative constructs or small molecules that target the ZZ domain of p62should block p62 function and inhibit support of MM cells and OCLformation by the marrow microenvironment.

Systematic biological studies of the inhibitors were performed as shownin FIG. 2.

Example 1 Compound XIELP1-106:N-(3,4-bis((4-fluorobenzyl)oxy)benzyl)-1-cyclohexylmethanaminehydrochloride

Clog RPMI8266 MM1S Internal ID Formula MW P (IC₅₀ μM) (IC₅₀ μM)XIELP1-106 C28H32ClFNO2 488.01 7.065 0.72 2.46

¹H NMR (DMSO-d₆): 9.16 (bs, 1H), 9.08 (bs, 1H), 7.92 (bs, 1H), 7.53-7.44(m, 5H), 7.24-7.19 (m, 4H), 7.10-7.05 (m, 2H), 5.13 (s, 4H), 4.01 (s,2H), 2.61 (bs, 2H), 1.75-1.60 (m, 5H), 1.22-1.10 (m, 4H), 0.93-0.84 (m,2H).

XIELP1-12b: N-(3,4-bis((4-fluorobenzyl)oxy)benzyl)cyclohexanamine

Clog RPMI8266 MM1S Internal ID Formula MW P (IC₅₀ μM) (IC₅₀ μM)XIELP1-12b C27H29F2NO2 437.52 6.75 1.18 1.77

¹H NMR (DMSO-d₆): 7.51-7.46 (m, 4H), 7.23-7.18 (m, 4H), 7.07-7.06 (m,1H), 6.98-6.96 (m, 1H), 6.84-6.82 (m, 1H), 5.09 (s, 2H), 5.07 (s, 2H),3.63 (s, 2H), 2.33-2.27 (m, 1H), 1.80-1.77 (m, 2H), 1.66-1.63 (m, 2H),1.54-1.50 (m, 1H), 1.16-0.99 (m, 5H).

XIELP1-17b: N-(3,4-bis((2,4-difluorobenzyl)oxy)benzyl)-1-cyclohexanamine

Clog RPMI8266 MM1S Internal ID Formula MW P (IC₅₀ μM) (IC₅₀ μM)XIELP1-17b C25H29F4NO2 487.53 7.35 0.94 0.84

¹H NMR (DMSO-d₆): 7.58-7.52 (m, 2H), 7.31-7.25 (m, 2H), 7.12-7.00 (m,4H), 6.86-6.84 (m, 1H), 5.10 (s, 2H), 5.08 (s, 2H), 3.58 (s, 2H),2.26-2.24 (m, 2H), 1.73-1.63 (m, 5H), 1.38-1.30 (m, 1H), 1.24-1.10 (m,3H), 0.87-0.79 (m, 2H).

XIELP1-20a: N-(3,4-bis((2,4-difluorobenzyl)oxy)benzyl)cyclohexanamine

Clog RPMI8266 MM1S Internal ID Formula MW P (IC₅₀ μM) (IC₅₀ μM)XIELP1-20a C27H27F4NO2 473.5 7.035 1.13 2.53

¹H NMR (DMSO-d₆): 7.58-7.52 (m, 2H), 7.31-7.25 (m, 2H), 7.15-7.07 (m,3H), 7.01-6.98 (m, 1H), 6.87-6.85 (m, 1H), 5.10 (s, 2H), 5.08 (s, 2H),3.63 (s, 2H), 2.32-2.27 (m, 1H), 1.80-1.54 (m, 6H), 1.17-1.12 (m, 3H),1.02-0.99 (m, 2H).

XIELP1-24b: 2-((3,4-bis((4-methylbenzyl)oxy)benzyl)amino)ethan-1-ol

MM1S U266 Internal Clog RPMI8266 (IC₅₀ (IC₅₀ ID Formula MW P (IC₅₀ μM)μM) μM) XIELP1- C25H29NO3 391.5 5.002 0.63 2.56 1.21 24b

¹H NMR (DMSO-d₆): 7.35-7.30 (m, 4H), 7.20-7.17 (m, 4H, 7.05-7.04 (m,1H), 6.96-6.94 (m, 1H, 6.81-6.79 (m, 1H), 5.04 (s, 2H), 5.03 (s, 2H),4.44 (t, J=5.2, 1H), 3.60 (s, 2H), 3.47-3.43 (m, 2H), 2.53-2.50 (m, 2H),2.31 (s, 3H), 2.30 (s, 3H), 1.98 (bs, 1H).

XIELP1-25b:N-(3,4-bis((4-methylbenzyl)oxy)benzyl)-1-cyclohexylmethanamine

MM1S U266 Internal Clog RPMI8266 (IC₅₀ (IC₅₀ ID Formula MW P (IC₅₀ μM)μM) μM) XIELP1- C30H37NO2 443.62 7.806 0.92 1.51 1.26 25b

¹H NMR (DMSO-d₆): 7.34-7.31 (m, 4H), 7.19-7.17 (m, 4H), 7.04-6.93 (m,2H), 6.79-6.77 (m, 1H), 5.04 (s, 2H), 5.03 (s, 2H), 3.56 (s, 2H), 2.31(s, 6H), 2.25 (d, J=6.8, 2H), 1.73-1.63 (m, 5H), 1.38-1.10 (m, 4H),0.88-0.81 (m, 2H).

XIELP1-51: N-(3,4-bis((4-methylbenzyl)oxy)benzyl)cyclohexanamine

Clog RPMI8266 MM1S Internal ID Formula MW P (IC₅₀ μM) (IC₅₀ μM)XIELP1-51 C29H35NO2 429.59 7.491 1.83 5.36

¹H NMR (DMSO-d₆): 7.34-7.30 (m, 4H), 7.19-7.17 (m, 4H), 7.05-6.93 (m,2H), 6.81-6.78 (m, 1H), 5.05 (s, 2H), 5.03 (s, 2H), 3.61 (s, 2H),2.28-2.34 (m, 5H), 1.20-0.96 (m, 5H).

XIELP1-58:N-(3,5-bis((2,4-difluorobenzyl)oxy)benzyl)-1-cyclohexylmethanamine

Clog RPMI8266 MM1S Internal ID Formula MW P (IC₅₀ μM) (IC₅₀ μM)XIELP1-58 C28H29F4NO2 487.53 7.35 1.40 2.50

¹H NMR (DMSO-d₆): 7.64-7.58 (m, 2H), 7.33-7.27 (m, 2H), 7.15-7.10 (m,2H), 6.63-6.62 (m, 2H), 6.56-6.55 (m, 1H), 5.08 (s, 4H), 3.61 (s, 2H),2.27 (d, J=6.4, 2H), 1.74-1.60 (m, 5H), 1.39-1.22 (m, 1H), 1.23-1.10 (m,3H), 0.88-0.82 (m, 2H).

XIELP1-60: N-(3,5-bis((2,4-difluorobenzyl)oxy)benzyl)cyclohexanamine

Clog RPMI8266 MM1S Internal ID Formula MW P (IC₅₀ μM) (IC₅₀ μM)XIELP1-60 C27H27F4NO2 473.5 7.035 1.33 2.51

¹H NMR (DMSO-d₆): 7.64-7.58 (m, 2H), 7.33-7.28 (m, 2H), 7.15-7.13 (m,2H), 6.63-6.55 (s, 3H), 5.08 (s, 4H), 3.66 (s, 2H), 2.33-2.28 (m, 1H),1.81-1.54 (m, 5H), 1.14-1.00 (m, 5H).

The therapeutic efficacy and tolerability of compound XIELP1-106 (alsoreferred to as 10b) was evaluated in the treatment of subcutaneousRPMI-8226 human multiple myeloma xenograft model. The tumor volumes inthe two groups at different time points are shown FIG. 3. The mean tumorvolume of the XIELP1-106 vehicle control group reached 1,452 mm3 on day39 after tumor implantation. The treatment of XIELP1-106 (60 mg/kg, ip,q3d×4 wks) produced a strong antitumor activity with T/C value of 23%(p=0.001).

The median survival time (MST) of the vehicle treated mice was 42 days(FIG. 4). In XIELP1-106 group, no animal death occurred until the end ofthe study (60 days post tumor inoculation).

Regarding the safety profile, the mice treated with XIELP1-106 at thegiven dose level yielded moderate body weight loss of 11.2% after 3doses, and then developed the tolerability to the treatment during therest of the period. No other gross clinical abnormality was observed. Insummary, XIELP1-106 at the given dose level demonstrated a strongantitumor efficacy in the treatment of RPMI-8226 human multiple myelomaxenograft model in this study, and significantly prolonged the survivaltime of the subcutaneous RPMI-8226 tumor-bearing mice.

In view of the many possible embodiments to which the principles of thedisclosed compounds, compositions and methods may be applied, it shouldbe recognized that the illustrated embodiments are only preferredexamples of the invention and should not be taken as limiting the scopeof the invention.

What is claimed is:
 1. A compound, or a pharmaceutically acceptable saltthereof, having a formula I of:

wherein Ar is benzenetriyl; R¹ has a structure of: CH₂—X—(CH₂)_(m)—R¹¹,wherein X is NH or O, m is 0 to 6, and R¹¹ is optionally-substitutedcycloalkyl; each R² is the same or different and has a structure of:

wherein Z is —O—; Z¹ is (—CH₂—)_(m); Cy is phenyl; and each R⁴ is thesame or different and is selected from hydroxy, halogen, substituted orunsubstituted alkoxy, substituted or unsubstituted alkyl, or amino; andc is 1 to 5; and each R³ is the same or different and is selected fromhydroxy, halogen, substituted or unsubstituted alkoxy, substituted orunsubstituted alkyl, amino, substituted or unsubstituted cycloalkyl,substituted or unsubstituted heterocycloalkyl, substituted orunsubstituted aryl, or a nitro, wherein a is 2, and b is
 0. 2. Thecompound of claim 1, wherein a first R² group is in a meta position onthe Ar ring relative to the R¹ group, and a second R² group is in a paraposition on the Ar ring relative to the R¹ group.
 3. The compound ofclaim 1, wherein R⁴ is —F, —Cl, —OCH₃, —OH, —CH₃ or —NH₂.
 4. Thecompound of claim 1, wherein c is 1 and R⁴ is 4-fluoro, or 4-methyl. 5.The compound of claim 1, wherein c is 2 and R⁴ is 2, 4-difluoro.
 6. Thecompound of claim 1, wherein R¹¹ is

and m is 0 or
 1. 7. The compound of claim 1, wherein each R² group hasthe same structure.
 8. The compound of claim 1, having a structure of:

wherein each of R⁷, R⁸, and R⁹ are the same or different and areselected from —F, —Cl, —OCH₃, —OH, —CH₃, or —NH₂; d is 0; e is 1 to 5; fis 1 to 5; m is 0 or 1; and R¹¹ is optionally-substituted cycloalkyl. 9.The compound of claim 1, having a structure of:

wherein each of R⁷, R⁸, and R⁹ are the same or different and areselected from —F, —Cl, —OCH₃, —OH, —CH₃, or —NH₂; d is 0 to 3; e is 1 to5; f is 1 to 5; m is 0 or 1; and R¹¹ is optionally-substitutedcycloalkyl.
 10. The compound of claim 8, wherein R¹¹ is


11. The compound of claim 1, selected from:


12. A compound, or a pharmaceutically acceptable salt thereof, having aformula I of:

wherein Ar is benzenetriyl; R¹ has a structure of:

wherein W is —CH₂—; X is —NR⁵—, wherein R⁵ is H or an alkyl, or —O—; andY is

each R² is the same or different and has a structure of:

wherein Z is —O—; Z¹ is (—CH₂—)_(m) wherein m is 1; Cy is phenyl; andeach R⁴ is the same or different and is selected from hydroxy, halogen,substituted or unsubstituted alkoxy, substituted or unsubstituted alkyl,or amino; and c is 0 to 5; and each R³ is the same or different and isselected from hydroxy, halogen, substituted or unsubstituted alkoxy,substituted or unsubstituted alkyl, amino, substituted or unsubstitutedcycloalkyl, substituted or unsubstituted heterocycloalkyl, substitutedor unsubstituted aryl, or a nitro, wherein a is 2 to 5, and b is 0 to 3.13. The compound of claim 12, wherein X is —NR⁵—, wherein R⁵ is H. 14.The compound of claim 1, wherein the compound is:


15. The compound of claim 12, wherein a is 2 and b is
 0. 16. Thecompound of claim 12, wherein R⁴ is —F, —Cl, —OCH₃, —OH, —CH₃ or —NH₂.17. The compound of claim 12, wherein c is 1 and R⁴ is 4-fluoro, or4-methyl.
 18. The compound of claim 12, wherein c is 2 and R⁴ is 2,4-difluoro.